JPH0548421B2 - - Google Patents

Description

[Detailed Description of the Invention] [Field of Application of the Invention] The present invention mainly relates to a two-dimensional electrophoresis device that enables simultaneous multi-component analysis of proteins, and particularly to a two-dimensional electrophoresis device that has a compact and functional electrophoresis tank. Regarding.

[Background of the invention]

In conventional horizontal electrophoresis devices, the electrolytic solution tank and the sample electrophoresis method are at right angles (Hirokawa Shoten "Latest Electrophoresis Method" p. 440) When electrophoresing multiple samples at the same time, the electrophoresis support must be cooled. They are arranged in parallel on a board, with both ends being positive and maintaining liquid contact with the catholyte. Two-dimensional electrophoresis involves the operation of moving a one-dimensional support, which is insulated and isolated from a two-dimensional support, to a predetermined position on the two-dimensional support at the end of electrophoresis. In order to automate this device, it is desirable to perform mechanical alignment with minimal movement.
It is essential that the device be as compact as possible.

However, such a compact device has not been proposed so far.

[Purpose of the invention]

An object of the present invention is to provide a compact automatic two-dimensional electrophoresis apparatus.

[Summary of the invention]

It is desirable for an automatic two-dimensional electrophoresis device to transfer this support onto a two-dimensional electrophoresis support in the simplest and least amount of movement at the end of one-dimensional electrophoresis. To do this, each one-dimensional support must be located opposite the two-dimensional support.
Furthermore, it is necessary to make the device as compact as possible, and the present invention has been conceived as a structure that satisfies the above conditions.

[Embodiments of the invention]

An embodiment of the present invention will be described below.

Figure 1 is a front view of a horizontal two-dimensional electrophoresis device, in which one-dimensional electrophoresis gel (for example, polyacrylamide for isoelectric focusing) is bonded and immobilized on one side to a substrate 4 and the other side is open. gel) 1
are placed on the cooling plate 3, and the substrates 4 are arranged in a row on the cooling plate 3. The cooling plate is a highly insulating and highly thermally conductive plate (e.g. SiC plate)
With a structure in which a Peltier element is closely attached to the bottom of the
A current is passed through the Peltier element to cool it (wiring to the Peltier element is not shown). Both ends of the gel for one-dimensional migration are oriented perpendicularly to the support to maintain liquid contact with the positive and negative electrolytes in the electrolyte baths 5 and 6, which are placed parallel to each other with a cooling plate in between. A liquid junction material (for example, paper) 9 is set. One electrode 10, 11 is provided in each of the electrolyte baths, and the structure is such that a voltage is applied to each terminal. 7 is a sample filling hole.

At the end of one-dimensional electrophoresis, the parts of the substrate exposed at both ends of this gel are sandwiched between supports (not shown), and the two-dimensional electrophoresis gel 2 (for example, a polyacrylamide concentration gradient gel with low It is filled in the groove 8 previously provided on the concentration side) as shown in FIG. A cooling plate 3' is also provided under the two-dimensional migration gel 2 and under the same substrate 4' as described above.

Two or more sets of the gel for one-dimensional migration and the gel for two-dimensional migration as described above are arranged laterally in the figure, and up to about 30 sets can be moved simultaneously by the gel moving device described later. However, it is preferable to move 5 to 20 sets at a time.

FIG. 3 is a diagram showing an example of a gel movement mechanism in the present invention. In the figure, a spring 4 is mounted on an arm 23 on which a substrate 4 on which a gel 1 for one-dimensional migration is mounted is rotated.
4, the front and rear of the board 4 are sandwiched and fixed. arm 2
The shaft 46 is inserted into the rotation hole 45 of No. 3, and the shaft 46 and the arm 23 are connected via a pin 47. A groove 48 of the arm 23 into which a pin 47 attached to the shaft 46 is inserted is an elongated hole in the direction of rotation shown by an arrow 49, and the shaft 46 and the arm 23 are coupled with play in the direction of rotation. A worm wheel 50 is attached to the other end of the shaft 46, and a motor 51
A dog 53 that controls the motor 51 during rotation is attached to the worm wheel 50 fixed to the shaft of the worm wheel 50.
The structure is such that the motor 51 is controlled by contacting with the motor 51. That is, electrophoresis of the gel 1 for one-dimensional migration is performed at the position of the limit switch 52, and then two-dimensional electrophoresis is performed at the position of the limit switch 43 as the rotating shaft 46 rotates.

A two-dimensional migration gel 2 is placed on the opposite side of the one-dimensional migration gel 1. After the one-dimensional migration gel 1 contacts and overlaps the groove 8 in the two-dimensional migration gel 2, a voltage is applied to both ends of the two-dimensional migration gel 2 to perform electrophoresis. Gel 1 for one-dimensional migration
A spring 58 is mounted on the arm 23 for rotational positioning. This spring 58
When it rotates toward the gel 2 for two-dimensional migration, it hits the fixing plate 59, thereby reducing the contact pressure of the gel 1 for one-dimensional migration against the gel 2 for two-dimensional migration.
If this contact pressure is large, the gel will be damaged, making electrophoresis impossible. The spring 58 is movable in the direction of the arrow 60 so that the contact pressure can be adjusted. Further, the position of the control limit switch 43 of the motor 51 is adjusted so that the motor 51 is stopped within the range of play in the direction of the arrow 49 of the arm 23.

Although one electrophoresis device has been described above, in reality, the shaft 46 is extended in the axial direction and a plurality of arms 23 are arranged, so that a plurality of electrophoresis gels can be moved simultaneously. .

FIG. 4 shows another embodiment of the invention. The figure is 1
Only one part for dimensional electrophoresis is shown. When the cooling plate is larger than the substrate, when the liquid junction material is deflected, the liquid junction material and the cooling plate come into contact. Then, the electrolytic solution spreads on the cooling plate, causing electrical leakage, which may prevent uniform current from flowing through the gel and reduce the reproducibility of electrophoresis.

Therefore, both ends of the cooling plate 3 are tilted as shown in FIG. 4, and the liquid junction material 9 is similarly tilted toward the electrolyte tanks 5 and 6. The angle of inclination of the cooling plate 3 is preferably about 10 to 35 degrees in order to facilitate drainage even if the electrolyte is spilled. The angle of inclination of the liquid junction material 9 is 5~
The angle is preferably about 30 degrees. If it is less than 5 degrees, deflection may occur, and if it exceeds 30 degrees, the contact between the gel and the liquid junction material will become unstable.

〔Effect of the invention〕

According to the present invention, it is possible to arrange both the one-dimensional and two-dimensional electrolyte baths and the cooling plate in parallel, making the entire system compact and contributing to effective use of space.

[Brief explanation of the drawing]

FIG. 1 is a front view of an embodiment of the two-dimensional electrophoresis device of the present invention, FIG. 2 is a sketch of the electrophoresis support, FIG. 3 is a sketch of the gel transfer mechanism, and FIG. FIG. 6 is a partial sketch of another embodiment. 1... Gel for one-dimensional migration, 2... Gel for two-dimensional migration, 3, 3'... Cooling plate, 4, 4'... Substrate,
5, 5'...Anolyte electrolyte tank, 6,6'...Cathode electrolyte tank, 7...Sample filling hole, 8...Groove, 9,9'
... Liquid junction material, 10 ... Electrode, 11 ... Electrode.

Claims (1)

[Scope of Claims] 1. In a horizontal two-dimensional electrophoresis device, a plurality of sets of one-dimensional electrophoresis gels and corresponding two-dimensional electrophoresis gels are arranged, and the plurality of one-dimensional electrophoresis gels The gels for one-dimensional electrophoresis are arranged substantially on a straight line at predetermined intervals in the direction, and the positive and cathode electrolyte baths for one-dimensional electrophoresis are respectively arranged almost parallel to this straight line, and each gel for one-dimensional electrophoresis is It is connected to the positive electrolytic solution tank for one-dimensional electrophoresis and the electrolytic solution tank for cathode, respectively, by a liquid junction material extending substantially perpendicular to the electrophoresis direction at both ends thereof, and is connected to the positive electrolytic solution tank for two-dimensional electrophoresis,
The cathode electrolyte tank is arranged substantially parallel to the straight line, the plurality of gels for two-dimensional migration are arranged in a row substantially parallel to the straight line, and each of the plurality of gels for two-dimensional migration is arranged substantially parallel to the straight line. An electrophoresis device characterized in that both ends perpendicular to the direction in which the gel for two-dimensional electrophoresis is arranged are connected to the positive and cathode electrolytic solution tanks for two-dimensional electrophoresis by liquid junction materials, respectively. 2. The electrophoresis apparatus according to claim 1, wherein there is one positive and one cathode electrolytic solution tank for one-dimensional electrophoresis. 3. According to claim 2, each of the gels for one-dimensional electrophoresis is installed between the anode electrolyte tank and the cathode electrolyte tank for one-dimensional electrophoresis. Electrophoresis apparatus described. 4 positive and negative electrolyte baths for one-dimensional electrophoresis;
A plurality of cooling plates each carrying a plurality of positive and negative electrolyte baths for the two-dimensional electrophoresis and a plurality of gels for the one-dimensional electrophoresis formed on the substrate are connected to the two-dimensional electrophoresis bath formed on the substrate. 4. The electrophoresis device according to claim 1, wherein the electrophoresis device is arranged substantially parallel to a single cooling plate on which a plurality of gels for electrophoresis are mounted.